NUCLEOSIDE TRIPHOSPHATES WITH MODIFIED PHOSPHATE CHAINS, AND METHODS OF SYNTHESIZING THE SAME
In some examples, a nucleoside triphosphate analogue may include a sugar, a nucleobase coupled to the sugar, a triphosphate group coupled to the sugar, a heteroatom coupled to an alpha phosphate of the triphosphate group, and a first substituent coupled to the heteroatom. The heteroatom may be selected from the group consisting of oxygen, nitrogen, and carbon. The first substituent may include at least one of an alkyl chain or a polymer.
1 . A nucleoside triphosphate analogue, comprising:
a sugar;
a nucleobase coupled to the sugar;
a triphosphate group coupled to the sugar;
a heteroatom coupled to an alpha phosphate of the triphosphate group; and
a first substituent coupled to the heteroatom.
2 . The nucleoside triphosphate analogue of claim 1 , wherein the heteroatom is selected from the group consisting of oxygen, nitrogen, and carbon.
3 . The nucleoside triphosphate analogue of claim 1 , wherein the first substituent comprises at least one of a fluorescent dye, an alkyl chain, and a polymer.
4 . The nucleoside triphosphate analogue of claim 3 , wherein the polymer comprises at least one of a synthetic peptide and a synthetic polymer.
5 . The nucleoside triphosphate analogue of claim 4 , wherein the synthetic polymer comprises an ethylene glycol polymer.
6 . The nucleoside triphosphate analogue of claim 3 , wherein the polymer comprises about 1 to about 50 repeating units.
7 . The nucleoside triphosphate analogue of claim 1 , wherein the first substituent comprises a functional group.
8 . The nucleoside triphosphate analogue of claim 7 , wherein the functional group comprises at least one of an amine, an azide, a carboxylic acid, a thiol, a tetrazine, a cyclooctyne, and an alkyne.
9 . The nucleoside triphosphate analogue of claim 1 , wherein the sugar comprises a natural ribose or deoxyribose.
10 . The nucleoside triphosphate analogue of claim 1 , wherein the nucleoside triphosphate analogue further comprises a second substituent coupled to the sugar via a 3′ carbon of the sugar.
11 . The nucleoside triphosphate analogue of claim 1 , wherein the second substituent comprises a protecting group.
12 . The nucleoside triphosphate analogue of claim 11 , wherein the protecting group comprises at least one of a vinyl group, an allyloxy methyl group, and an azido methyl group.
13 . The nucleoside triphosphate analogue of claim 1 , wherein the nucleoside triphosphate analogue comprises a third substituent coupled to the sugar via a 2′ carbon of the sugar.
14 . The nucleoside triphosphate analogue of claim 13 , wherein the third substituent comprises at least one of a fluorine, a methoxy group, and an ethoxy group.
15 . The nucleoside triphosphate analogue of claim 1 , wherein the nucleoside triphosphate analogue further comprises a fourth substituent coupled between a beta phosphate and a gamma phosphate of the triphosphate group.
16 . The nucleoside triphosphate analogue of claim 15 , wherein the fourth substituent comprises at least one of an oxygen atom, a nitrogen atom, a methylene, a fluoromethylene, a difluoromethylene, a sulfur atom, and a selenium atom.
17 . The nucleoside triphosphate analogue of claim 1 , wherein the nucleobase is a non-naturally occurring nucleobase.
18 . A method of synthesizing a nucleoside triphosphate analogue, the method comprising:
coupling a phosphorylating agent to a sugar of a nucleoside, the nucleoside comprising the sugar and a nucleobase, the phosphorylating agent comprising a protecting group and a substituent;
oxidizing the coupled phosphorylating agent;
removing the protecting group from the oxidized, coupled phosphorylating agent to form a monophosphate diester comprising the substituent;
activating the monophosphate diester; and
reacting the activated monophosphate diester with a pyrophosphate to form a triphosphate group comprising the substituent.
19 . The method of claim 18 , wherein the sugar comprises a protected 3′-O group.
20 - 32 . (canceled)
33 . A method of synthesizing a nucleoside triphosphate analogue, the method comprising:
activating a monophosphate group of a nucleotide, the nucleotide comprising a sugar, a nucleobase, and the monophosphate group;
reacting the activated monophosphate group with an amidation agent comprising a substituent in an amidation reaction to form an amidated monophosphate group comprising the substituent;
activating the amidated monophosphate group; and
reacting the activated, amidated monophosphate group with a pyrophosphate to form a triphosphate group comprising the substituent.
34 - 85 . (canceled)